Antibodies, Proteins, Kits and Reagents for Life Science

307 566 ₸ Product size

100 µl 307 566 ₸ 10 µl 90 460 ₸

Order now and get it on Tuesday March 09, 2021

Anti-BRN3A antibody [EPR23257-285] (ab245230)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23257-285] to BRN3A
Suitable for: IHC-Fr, IP, Flow Cyt, IHC-P, WB
Reacts with: Mouse, Rat, Human, Recombinant fragment

Product name

Anti-BRN3A antibody [EPR23257-285]
See all BRN3A primary antibodies
Description

Rabbit monoclonal [EPR23257-285] to BRN3A
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-Fr	Mouse
IHC-P	Mouse Rat
IP	Mouse
WB	Mouse Human Recombinant fragment

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: MOLT4 whole cell lysate. Mouse E14.5, mouse E18 and rat E18 brain tissue lysate. His-tagged Mouse POU4F1 recombinant protein.Flow Cyt: MOLT4 cells. IHC-P: Mouse E14.5 cochleovestibular ganglion and colliculus. Rat E14.5 dorsal root ganglia.IHC-Fr: Mouse retina tissue. Mouse E14.5 embryo. IP: Mouse E14.5 brain tissue lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23257-285
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A antibody [EPR23257-285] (ab245230)

Immunohistochemical analysis of paraffin-embedded mouse E14.5 tissue labeling BRN3A with ab245230 at 1/2000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on cochleovestibular ganglion and colliculus of mouse E14.5 (PMID: 24709358). The section was incubated with ab245230 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-BRN3A antibody [EPR23257-285] (ab245230)

All lanes : Anti-BRN3A antibody [EPR23257-285] (ab245230) at 1/1000 dilution

Lane 1 : MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate
Lane 2 : Daudi (human Burkitts lymphoma lymphoblast), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 43 kDa
Observed band size: 42 kDa
why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

This blot was developed using a higher sensitivity ECL substrate.

Negative control: Daudi (PMID: 20460523).

Exposure time: 3 minutes.
Flow Cytometry - Anti-BRN3A antibody [EPR23257-285] (ab245230)

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol-permeabilized Daudi (Human Burkitt's lymphoma lymphoblast (Left) / MOLT-4 (Human lymphoblastic leukemia T lymphoblast (Right) cells labelling BRN3A with ab245230 at 1/400 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control: Daudi (PMID: 20460523).
Western blot - Anti-BRN3A antibody [EPR23257-285] (ab245230)

All lanes : Anti-BRN3A antibody [EPR23257-285] (ab245230) at 1/1000 dilution

Lane 1 : Mouse E14.5 brain tissue lysate
Lane 2 : Mouse E18 brain tissue lysate
Lane 3 : Rat E18 brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 43 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

This blot was developed using a higher sensitivity ECL substrate.

Exposure time: 3 minutes.
Immunohistochemistry (Frozen sections) - Anti-BRN3A antibody [EPR23257-285] (ab245230)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse retina tissue labeling BRN3A with ab245230 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Nuclear staining on ganglion cell layer of mouse retina is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunoprecipitation - Anti-BRN3A antibody [EPR23257-285] (ab245230)

BRN3A was immunoprecipitated from 0.35 mg mouse E14.5 brain tissue lysate with ab245230 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab245230 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse E14.5 brain tissue lysate 10µg.

Lane 2: ab245230 IP in Mouse E14.5 brain tissue lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245230 in mouse E14.5 brain tissue lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.

This blot was developed using a higher sensitivity ECL substrate.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A antibody [EPR23257-285] (ab245230)

Immunohistochemical analysis of paraffin-embedded rat E14.5 tissue labeling BRN3A with ab245230 at 1/2000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on dorsal root ganglia of rat E14.5 (PMID: 8341591). The section was incubated with ab245230 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Frozen sections) - Anti-BRN3A antibody [EPR23257-285] (ab245230)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse E14.5 embryo tissue labeling BRN3A with ab245230 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Nuclear staining on mouse E14.5 embryonic brain is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Western blot - Anti-BRN3A antibody [EPR23257-285] (ab245230)

All lanes : Anti-BRN3A antibody [EPR23257-285] (ab245230) at 1/1000 dilution

Lane 1 : His-tagged Mouse POU4F1 recombinant protein (aa2-100-GGGS linked aa2-100), 10ng
Lane 2 : His-tagged Mouse POU4F2 recombinant protein (aa91-261), 20ng
Lane 3 : His-tagged Mouse POU4F3 recombinant protein (aa2-100), 20ng

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 43 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

ab245230 has a low affinity with mouse POU4F2 recombinant protein.

Exposure time: 15 seconds.
Anti-BRN3A antibody [EPR23257-285] (ab245230)

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